Figure 6. Silencing of IκBζ expression in human Vγ9Vδ2 T cells attenuates IFN-γ production in response to IL-12 and IL-18.

Ex vivo-expanded Vγ9Vδ2 T cells were transfected with siRNA targeting IκBζ or control siRNA followed by 16-hour treatment of IL-12 and IL-18. Zombie violet positive cells were gated-out as dead cells. (A) Representative contour plots of IκBζ-positive cells gated on CD3⁺ Vδ2 TCR⁺ cells. Among three different siRNAs targeting IκBζ, one (s34643) silenced cytokine-induced IκBζ efficiently, and which was used for knocking down IκBζ in further experiments. (B) Representative contour plots and percentage of BTLA-positive cells by CD3⁺Vδ2 TCR⁺ cells. (C-F) Representative histogram (left) and gMFI (right) of CD25 (C), granzyme B (D), ICAM-1 (E) and IFN-γ (F) by CD3⁺Vδ2 TCR⁺ cells. The data were obtained from three independent experiments each using Vγ9Vδ2 T cells from different donors. Bars represent the mean and SEM, *P < 0.05, two-tailed paired Student t test.